Cryobiology最新文献

{"title":"Cryopreservation of hormonally induced sperm of the common toad (Bufo bufo) with a cushion centrifugation step: a preliminary study","authors":"Natalia V. Shishova,&nbsp;Svetlana A. Kaurova,&nbsp;Victor K. Uteshev","doi":"10.1016/j.cryobiol.2026.105594","DOIUrl":"10.1016/j.cryobiol.2026.105594","url":null,"abstract":"<div><div>One of the most important methods for preserving amphibian biodiversity is sperm cryobanking, in particular cryopreservation of spermic urine obtained by hormonal induction of spermiation. However, for bufonids, cryopreserved sperm obtained by this method has low fertilizing ability, which hampers its use in cryobanking and artificial breeding programs for endangered species. We hypothesized that the key cause of this problem is the low concentration of sperm in the spermic urine of toads. It is possible to increase sperm concentration by centrifugation. The purpose of this work is to study the resistance of common toad (<em>Bufo bufo</em>) sperm to centrifugation and the effect of centrifugation on sperm cryoresistance. The resulting samples were subjected to noncushioned ( × 200 g, 5 min and × 500 g, 5 min) and cushioned ( × 500 g, 5 min, with underlayering of a cushioned solution: Niu-Twitty medium + 10% bovine serum albumin (BSA) + 5% sucrose). A comparison was also made of programmable freezing (2 °C/min), freezing in nitrogen vapor (7-9 °C/min), and three thawing methods (ice bath, air at 18 °C, and water bath at 40 °C). It was found that noncushioned centrifugation did not reduce sperm motility but reduced its fertilizing ability. Cushioned centrifugation did not have a negative effect on the fertilizing ability of common toad sperm. The fertilizing ability of sperm frozen after cushioned centrifugation was 39 ± 10%, which significantly exceeded the fertilizing ability of sperm frozen without centrifugation (&lt;1%) and after noncushioned centrifugation (4%). Programmable freezing followed by slow thawing in air resulted in better sperm survival compared with more rapid freezing and thawing.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105594"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146212490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryopreservation of Drosophila melanogaster embryonic nuclei in a Dimethyl sulfoxide (ME2SO)-free solution","authors":"Troy Louwagie ,&nbsp;Lindsey G. O’Brien ,&nbsp;Daryl M. Gohl ,&nbsp;Allison Hubel","doi":"10.1016/j.cryobiol.2025.105561","DOIUrl":"10.1016/j.cryobiol.2025.105561","url":null,"abstract":"<div><div>Drosophila melanogaster is a well-known organism for biological research. With more than 150,000 live stocks maintained, there is an urgent need for cryopreservation of Drosophila stocks to limit maintenance costs and genetic drift. Two methods have been described for freezing Drosophila stocks: vitrification of embryos and of primordial germ cells. These methods show success in cryopreserving stocks, but the robustness and the applicability of these approaches in preserving a wide range of mutant stocks has not been established. Here, we describe a method for cryopreserving isolated embryonic nuclei using a slow-cooling method. Using a differential evolution algorithm, we identified five different formulations that show <span><math><mo>&gt;</mo></math></span>80% recovery and used Raman spectroscopy to image nuclei at low temperatures. Finally, we demonstrate that cryopreserved nuclei retain biological function through an induced <em>hsp70</em> transcriptional response. This work represents steps towards cryopreserving nuclei with the ultimate goal of regenerating stocks by embryonic nuclear transplantation.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105561"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Age leaves a molecular mark: altered A‐Kinase Anchor Protein 4 (proAKAP4) abundance and sperm cell quality drive fertility decline in Bali bulls (Bos sondaicus)","authors":"Irma Octaria Sari ,&nbsp;Berlin Pandapotan Pardede ,&nbsp;Mokhamad Fakhrul Ulum ,&nbsp;Herdis ,&nbsp;Bambang Purwantara ,&nbsp;Erdogan Memili","doi":"10.1016/j.cryobiol.2026.105592","DOIUrl":"10.1016/j.cryobiol.2026.105592","url":null,"abstract":"<div><div>Reproductive performance in bulls is strongly influenced by age-related factors, which can lead to functional and molecular deterioration of sperm even when conventional semen parameters remain within acceptable ranges. This study aimed to evaluate the impact of the age factor on sperm quality and fertility in Bali bulls (<em>Bos sondaicus</em>), with particular emphasis on proAKAP4, the precursor of A-Kinase Anchor Protein 4, an essential regulator of flagellar structure and motility. Seventeen Bali bulls routinely used in artificial insemination (AI) programs were classified into two age categories: mature bulls (≤9 years; n = 8) and older-age bulls (≥11 years; n = 9). Post-thaw semen samples were evaluated for progressive and total motility, sperm kinematics, viability, morphological abnormalities, DNA fragmentation, and protamine deficiency. ProAKAP4 concentrations were quantified using ELISA. Field fertility outcomes were derived from &gt;10,000 AI records collected over two years. Compared with mature bulls, older-age bulls exhibited significantly lower (P &lt; 0.05) motility, viability, and kinematic parameters, alongside higher rates of morphological abnormalities, DNA fragmentation, and protamine deficiency. ProAKAP4 abundance was markedly reduced in older-age bulls and closely mirrored their lower fertility performance. ProAKAP4 levels were positively correlated with motility, viability, and fertility rate, and negatively correlated with morphological and nuclear defects (P &lt; 0.05). In conclusion, the age factor plays a critical role in determining sperm quality and fertility potential in Bali bulls, with proAKAP4 emerging as a sensitive molecular biomarker of sperm function and field fertility. Integrating proAKAP4 quantification into routine semen evaluation may enhance sire selection accuracy and overall reproductive efficiency in AI programs. Considering the bull's age is therefore essential, as it directly influences proAKAP4 abundance, sperm functional integrity, and fertility outcomes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105592"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146156437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Similar developmental outcomes of salamander larvae produced with fresh and cryopreserved sperm: implications for amphibian conservation","authors":"Shaina Lampert ,&nbsp;Isabella Burger ,&nbsp;Devin Chen ,&nbsp;Andy Kouba ,&nbsp;Carrie Kouba","doi":"10.1016/j.cryobiol.2026.105593","DOIUrl":"10.1016/j.cryobiol.2026.105593","url":null,"abstract":"<div><div>Sperm cryopreservation is an important tool for the genetic management of threatened amphibian species within conservation breeding programs. Yet, little is known about the growth and development of offspring generated using frozen-thawed sperm compared to those produced using fresh sperm. The objective of this study was to compare the fertilization rate, embryo development, morphology, and survival of tiger salamander (<em>Ambystoma tigrinum</em>) offspring generated using either fresh or frozen-thawed sperm. Captive male and female <em>A. tigrinum</em> were administered exogenous hormones to stimulate gamete production and release. Eggs were collected from females and fertilized with either freshly collected sperm from captive males or frozen-thawed sperm maintained in the U.S. National Amphibian Genome Bank. Following <em>in-vitro</em> fertilization, egg fertilization rates and early embryo development were monitored through hatching. Larvae from each sperm type were then maintained under standardized rearing conditions and routinely monitored for total length, head width, survival, and development through metamorphosis. Results showed no difference in larval growth (<em>p</em> &gt; 0.42), survival rate (<em>p</em> = 0.72), or size at metamorphosis (<em>p</em> &gt; 0.26) between offspring generated from fresh or frozen-thawed sperm. These findings support sperm cryopreservation as an effective conservation tool for producing viable individuals in amphibian conservation programs.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105593"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146164595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In memoriam: James Lovelock (1919-2022) - A scientific life in cryobiology and beyond.","authors":"Sanaz Hemmatibardehshahi, Celina Phan, Jason P Acker","doi":"10.1016/j.cryobiol.2025.105547","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2025.105547","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"105547"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147431413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adding lycopene to the freezing media enhances the quality, antioxidant capacity, and fertilization ability of frozen-thawed Oure-type Tibetan sheep (Ovis aries) sperm","authors":"Yujie Tang ,&nbsp;Hong Wu ,&nbsp;Lidan Liu ,&nbsp;Yanbing Liu ,&nbsp;Haoqiang Qin ,&nbsp;Mingxia Li ,&nbsp;Deqing Yan ,&nbsp;Chengtu Zhang ,&nbsp;Jianmin Su","doi":"10.1016/j.cryobiol.2025.105578","DOIUrl":"10.1016/j.cryobiol.2025.105578","url":null,"abstract":"<div><div>Lycopene (LYC) is a plant-derived antioxidant that ameliorates oxidative and other stress-related damage in spermatozoa yet its inclusion in ovine freezing medium remains largely unexplored. In this study semen was collected from eight Oura-type Tibetan sheep with three ejaculates per male and cryopreserved in freezing medium supplemented with 0, 1, 2, 4, 8 and 16 μM LYC to identify the optimal concentration and evaluate its effects on structural integrity antioxidant status and fertilizing capacity. A dose of 2 μM LYC proved optimal markedly improving post-thaw survival and motion kinetics relative to the control (<em>P</em> &lt; 0.01). At both 25 °C and 4 °C this concentration prolonged survival and increased the proportion of live cells at each observation point. It also enhanced acrosome (<em>P</em> &lt; 0.01), head membrane (<em>P</em> &lt; 0.05) and tail membrane (<em>P</em> &lt; 0.01) integrity and elevated mitochondrial membrane potential (MMP) (<em>P</em> &lt; 0.05). Compared with the control the 2 μM LYC group showed lower reactive oxygen species (ROS) and malondialdehyde (MDA) levels (<em>P</em> &lt; 0.01) and higher superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) (<em>P</em> &lt; 0.05 and <em>P</em> &lt; 0.01 respectively). <em>In vitro</em> fertilization (IVF) trials revealed that 2 μM LYC increased cleavage (<em>P</em> &lt; 0.05) and blastocyst formation rates (<em>P</em> &lt; 0.01) without affecting blastocyst cell number (<em>P</em> &gt; 0.05). In a cervical artificial insemination (AI) programme pregnancy rates were 44 % for the 2 μM LYC group versus 28 % for the control. We conclude that adding 2 μM LYC to the freezing medium improves post-thaw quality counters oxidative stress and enhances the fertilizing capacity of Oura-type Tibetan sheep spermatozoa.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105578"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The application of artificial intelligence in cryopreservation: Technological advances and future challenges","authors":"Xingyue Lei ,&nbsp;Liqun He ,&nbsp;Gang Zhao","doi":"10.1016/j.cryobiol.2026.105589","DOIUrl":"10.1016/j.cryobiol.2026.105589","url":null,"abstract":"<div><div>This review systematically sorts out the latest research progress and application status of artificial intelligence (AI) technology in the field of cryopreservation, with a focus on its action mechanisms in aspects such as protocol optimization, damage prediction, and quality control in the cryopreservation of biological samples like cells, embryos, and tissues. From the perspectives of cross-scale modeling, dynamic process optimization, and prediction of structural and functional states, the article analyzes the advantages and challenges of AI in improving the efficiency of cryopreservation, reducing cell damage, and increasing the survival rate after resuscitation. It also summarizes the key application cases in the design of cryoprotectant (CPA) formulations, the control of the freezing or thawing process, and the monitoring of long-term stability. In view of the deficiencies of current AI models in sample diversity, mechanism analysis, and clinical translation, the review puts forward future development directions, including multi-modal data fusion, enhanced interpretability, and the construction of intelligent adaptive control systems. This research provides a reference for the intelligent development of cryopreservation technology and lays a theoretical foundation for subsequent clinical and industrial applications.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105589"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146124265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In Memoriam: Dr. Arthur W. Rowe","authors":"William A. Rowe,&nbsp;Chris Rowe Taitt","doi":"10.1016/j.cryobiol.2026.105587","DOIUrl":"10.1016/j.cryobiol.2026.105587","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105587"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicles as emerging bioactive modulators in sperm cryopreservation: Mechanisms and therapeutic potentials","authors":"Merve Deniz Tanrıkulu ,&nbsp;Mesut Çevik ,&nbsp;Mustafa Numan Bucak ,&nbsp;Alper Koçyiğit","doi":"10.1016/j.cryobiol.2025.105568","DOIUrl":"10.1016/j.cryobiol.2025.105568","url":null,"abstract":"<div><div>Sperm cryopreservation is a cornerstone of assisted reproduction and genetic conservation, yet the freeze-thaw process induces substantial structural and functional damage to spermatozoa. Recent progress in the field has positioned extracellular vesicles (EVs) as potential therapeutic tools, primarily because of their capacity to transport functional biomolecules that help counteract the cellular damage caused by cryopreservation. We then examine the types of cellular and molecular damages encountered during sperm cryopreservation and synthesize current evidence on the protective effects of EVs in this context. Particular attention is given to the mechanisms by which EVs may preserve sperm integrity, including membrane stabilization, antioxidant delivery, and modulation of signaling pathways. Despite encouraging findings, challenges remain, including the standardization of EVs isolation methods, optimal dosing, and delivery strategies. We also discuss the need for mechanism-of-action studies and multi-omics approaches to better understand EV-sperm interactions. Finally, future directions are outlined, including EVs engineering and clinical translation. Overall, EVs emerge as promising modulators in cryobiology, with potential to improve sperm preservation outcomes and fertility restoration strategies. This review offers an in-depth examination of EVs, covering the processes involved in their formation, various strategies employed for their isolation, and the techniques used to assess their properties. It also highlights their sources and functional significance in the male reproductive system.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105568"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryopreservation decreased lipopolysaccharide-induced immune response of endometrial stromal cells via inhibiting the expression of TRIF","authors":"Cuiting Yang ,&nbsp;Yan Zhang ,&nbsp;Chun Wang ,&nbsp;Lingxi Zhu ,&nbsp;Ming Zhang","doi":"10.1016/j.cryobiol.2025.105567","DOIUrl":"10.1016/j.cryobiol.2025.105567","url":null,"abstract":"<div><div>In the study, we investigated the effect of cryopreservation on the immune response of endometrial stromal cells (ESCs) to lipopolysaccharide (LPS). We exposed both frozen-thawed and untreated ESCs to LPS and measured cytokine levels as well as genes associated with the LPS-TLR4-cytokine signaling pathway. Results revealed that LPS induced significant upregulation of <em>IL-1β</em>, <em>IL-6</em>, <em>IL-8</em>, and <em>TNF-α</em> in vitro cultured untreated cells compared to frozen-thawed ones. Furthermore, mRNA levels related to the TRIF pathway, including <em>TRAM</em>, <em>TRAF-6</em>, and <em>IRF-7</em>, <em>NF-κB</em>, and <em>JNK</em> were significantly upregulated in untreated ESCs after LPS-stimulation but not in frozen-thawed cells. Moreover, transcription levels of TRIF signaling-related genes were notably lower in frozen-thawed cells. Additionally, TRIM69 overexpression rescued the cryopreservation-induced suppression of immune response. Overall, our findings suggest that cryopreservation attenuates the LPS-induced immune response in ESCs by inhibiting the TRIF pathway.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105567"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}